Fig. 1: LRPAP121-30S_V-specific CD8+ T-cell responses after vaccination. | Nature Communications

Fig. 1: LRPAP121-30S_V-specific CD8+ T-cell responses after vaccination.

From: TEIPP-vaccination in checkpoint-resistant non-small cell lung cancer: a first-in-human phase I/II dose-escalation study

Fig. 1

The presence of circulating LRPAP121-30-specific CD8+ T cells was determined by dual LRPAP121-30S- and LRPAP121-30V-dextramer staining using spectral flow cytometry after one round of in vitro expansion with the LPRPAP121-30V peptide. a, b Percentage of LRPAP121-30S_V-specific CD8+ T cells in the PBMC at baseline (BL) and after 1, 2 and 3 vaccinations (v1-v3) is provided for 23 patients across (a) all and (b) the different cohorts. c–e Hierarchically clustered heatmaps with the protein expression levels of indicated markers on CD8+ T cells non-specific (LRPAP-) or specific (LRPAP+ ) for LRPAP121-30, depicted (c) at baseline and after 1 (V1), 2 (V2) and 3 (V3) vaccinations, (d) and across the different cohorts, for 20 patients with a detectable LRPAP21-30S_V-specific CD8+ T-cell response, and (e) for ex vivo (d0) and in vitro expanded (d10) CD8+ T cells after 3 vaccinations (n = 2 patients from cohort 4). Marker expression is shown as z-score of median signal intensity per channel. Blue: low expression, red: high expression. f Percentage of single (SP; yellow), double (DP; green), triple (TP; orange), quadruple (Q4P; blue), and quintuple (Q5P; purple) cytokine-positive LRPAP121-30V-specific CD8+ T cells is depicted at baseline (BL) and after 1, 2 and 3 vaccinations (v1-v3) for 12 patients in which we could detect LRPAP121-30S_V-specific CD8+ T cells by dual dextramer analysis and had sufficient cells to perform additional intracellular cytokine staining. P-values versus baseline are indicated at the side of the bar. g LRPAP121-30S_V -specific CD8+ T cells were isolated from three patients after vaccination by dextramer-guided flow sorting, after which the LRPAP1 dextramer-positive T cell fractions were expanded and tested against WT, TAPKO and TAPKO/LRPAP1KO 518A2 melanoma cells (blue, red and green, respectively) as well as the LRPAP1-21-30S and -21-30V peptides (purple and orange, respectively). CCL4 (left) and IFNγ (right) production is determined by ELISA after overnight co-incubation of the T cells with the target cells. Data are represented as scatter plots with bars indicating mean and dots representing individual data points (a, b, g) or as stacked bars (f). Statistical analysis by unpaired non-parametric Kruskal-Wallis with Dunn’s multiple comparisons test (a, b, f, g). Source data are provided as a Source Data file.

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